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Journal of clinical microbiology2022; 61(1); e0123422; doi: 10.1128/jcm.01234-22

Protein Microarray-Guided Development of a Highly Sensitive and Specific Dipstick Assay for Glanders Serodiagnostics.

Abstract: Burkholderia mallei, the causative agent of glanders, is a clonal descendant of Burkholderia pseudomallei, the causative agent of melioidosis, which has lost its environmental reservoir and has a restricted host range. Despite limitations in terms of sensitivity and specificity, complement fixation is still the official diagnostic test for glanders. Therefore, new tools are needed for diagnostics and to study the B. mallei epidemiology. We recently developed a highly sensitive serodiagnostic microarray test for human melioidosis based on the multiplex detection of B. pseudomallei proteins. In this study, we modified our array tests by using anti-horse IgG conjugate and tested sera from B. mallei-infected horses (n = 30), negative controls (n = 39), and horses infected with other pathogens (n = 14). Our array results show a sensitivity of 96.7% (confidence interval [CI] 85.5 to 99.6%) and a specificity of 100.0% (CI, 95.4 to 100.0%). The reactivity pattern of the positive sera on our array test allowed us to identify a set of 12 highly reactive proteins of interest for glanders diagnosis. The B. mallei variants of the three best protein candidates were selected for the development of a novel dipstick assay. Our point-of-care test detected glanders cases in less than 15 min with a sensitivity of 90.0% (CI, 75.7 to 97.1%) and a specificity of 100.0% (CI, 95.4 to 100.0%). The microarray and dipstick can easily be adopted for the diagnosis of both B. mallei and B. pseudomallei infections in different animals. Future studies will show whether multiplex serological testing has the potential to differentiate between these pathogens.
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Publication Date: 2022-12-21 PubMed ID: 36541753PubMed Central: PMC9879090DOI: 10.1128/jcm.01234-22Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study focuses on refining diagnostic tests for glanders, a disease caused by Burkholderia mallei bacteria. Researchers have developed a highly sensitive and specific dipstick test that can accurately diagnose the disease in horses within 15 minutes using certain proteins identified from the bacteria.

Objective of the Study

  • This research aimed at developing a more precise and faster diagnostic method for glanders, a bacterial disease caused by Burkholderia mallei. It particularly pointed towards upgrading the existing diagnostic methods like complement fixation, which are suffering from limitations concerning sensitivity and specificity.

Progress of the Study

  • The research team initially developed a high-sensitivity microarray test for detecting a similar bacteria, B. pseudomallei, which causes melioidosis in humans.
  • They modified this test using an anti-horse IgG conjugate and then tested it on sera taken from B. mallei-infected horses, non-infected controls and horses infected with other pathogens.
  • This modified test displayed a high sensitivity and specificity: 96.7% sensitive and 100% specific.

Identifying Proteins for Diagnosis

  • The research team examined the positive sera’s reactivity layout in the array tests, enabling them to identify 12 highly reactive proteins crucial for diagnosing glanders.
  • The researchers selected the B. mallei variants of the top three protein candidates for developing an innovative dipstick assay.

Results of the Developed Dipstick Assay

  • The point-of-care dipstick test could identify glanders cases within less than 15 minutes.
  • With a 90.0% sensitivity and 100.0% specificity, it proved to be a highly successful diagnostic tool.

Implications and Future Work

  • The study concluded that both the microarray and the dipstick can be conveniently used to diagnose infections caused by B. mallei and B. pseudomallei in various animals.
  • Future research will be undertaken to discover if this multiplex serological testing can effectively differentiate between these two pathogens.

Cite This Article

APA
Wagner GE, Berner A, Lipp M, Kohler C, Assig K, Lichtenegger S, Saqib M, Müller E, Trinh TT, Gad AM, Söffing HH, Ehricht R, Laroucau K, Steinmetz I. (2022). Protein Microarray-Guided Development of a Highly Sensitive and Specific Dipstick Assay for Glanders Serodiagnostics. J Clin Microbiol, 61(1), e0123422. https://doi.org/10.1128/jcm.01234-22

Publication

Journal of clinical microbiology
ISSN: 1098-660X
NlmUniqueID: 7505564
Country: United States
Language: English
Volume: 61
Issue: 1
Pages: e0123422

Researcher Affiliations

Wagner, Gabriel E
  • Diagnostic & Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria.
Berner, Andreas
  • Diagnostic & Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria.
Lipp, Michaela
  • Diagnostic & Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria.
Kohler, Christian
  • Friedrich Loeffler Institute for Medical Microbiology, Greifswald, Germany.
Assig, Karoline
  • Diagnostic & Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria.
Lichtenegger, Sabine
  • Diagnostic & Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria.
Saqib, Muhammad
  • Department of Clinical Medicine and Surgery, University of Agriculture, Faisalabad, Pakistan.
Müller, Elke
  • InfectoGnostics Research Campus, Centre for Applied Research, Jena, Germany.
  • Leibniz-Institute of Photonic Technology (Leibniz-IPHT), Jena, Germany.
Trinh, Trung T
  • Institute of Microbiology and Biotechnology, Vietnam National University, Hanoi, Vietnam.
Gad, Anne-Marie
  • Senova Gesellschaft für Biowissenschaft und Technik mbH, Weimar, Germany.
Söffing, Hans Hermann
  • Senova Gesellschaft für Biowissenschaft und Technik mbH, Weimar, Germany.
Ehricht, Ralf
  • InfectoGnostics Research Campus, Centre for Applied Research, Jena, Germany.
  • Leibniz-Institute of Photonic Technology (Leibniz-IPHT), Jena, Germany.
  • Friedrich Schiller University Jena, Institute of Physical Chemistry, Jena, Germany.
Laroucau, Karine
  • Paris Est University, Anses, Animal Health Laboratory, Bacterial zoonosis UnitMaisons-Alfort Cedex, France.
Steinmetz, Ivo
  • Diagnostic & Research Institute of Hygiene, Microbiology and Environmental Medicine, Medical University of Graz, Graz, Austria.

MeSH Terms

  • Humans
  • Horses
  • Animals
  • Glanders / diagnosis
  • Melioidosis / diagnosis
  • Melioidosis / veterinary
  • Protein Array Analysis
  • Burkholderia mallei / genetics
  • Burkholderia pseudomallei

Conflict of Interest Statement

The authors A.-M.G. and H.H.S. declare a conflict of interest. A.-M.G. and H.H.S. are employees of Senova Gesellschaft für Biowissenschaft und Technik mbH, the company that manufactures the dipsticks. This does not affect the authors’ adherence to all the ASM policies on sharing data and materials. All other authors have declared that no competing interests exist.

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Citations

This article has been cited 9 times.
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